DRx brochure cell-based overview final v1 - DiscoverX Solutions for Drug Discovery · 2016. 10. 12. · Kinase targets have been extensively studied in biochemical assays using purified

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DiscoveRx Corporation42501 Albrae astreetFremont, CA 94538United States

To place an order:t | 510.979.1415f | 510.979.1650e | [email protected] | 866.448.4864www.discoverx.com

PathHunter™ Cell-Based Kinase Assays

Your Key To Elucidating Novel Kinase Therapeutics

DRX_KINASE_OVERVIEW_V2_0310

©2010 DiscoveRx Corporation. All Rights Reserved. DiscoveRx logo, DiscoveRx, PathHunter and HitHunter are trademarks of DiscoveRx Corporation.

European Regional Headquarters

DiscoveRx Corporation Ltd. (United Kingdom) Faraday Wharf, Holt Street Birmingham Science Park Aston Birmingham, B7 4BB To place an order: t | +44.121.260.6142 f | +44.121.260.6143e | [email protected]

PathHunter™ Cell-Based Kinase AssaysUnique, whole cell assay platform for screening and profiling of

Tyrosine Kinase Activity at the Receptor and Beyond the Receptor

Call 1.866.448.4864 to place your order today!For an updated list of available targets, visit www.discoverx.com/kinases

Kinases are a large and complex class of highly druggable target proteins that are inherent to most signaling pathways. DiscoveRx Offers novel, target-specific cell-based assays to study a variety of kinase targets at the receptor and generic pathway relevant assays for intractable targets. Target-specific assays offer fewer false positives, more relevant pharmacology and lesser chance of compound interference, while the pathway assays from DiscoveRx offer broad applicability and downstream read-outs.

Why Cell-Based Assays for Kinases?

Kinase targets have been extensively studied in biochemical assays using purified protein fragments for the kinase and the substrate. Many drugs have been identified using in-vitro biochemical assays, such as enzyme activity and receptor binding. However, there is an increasing need to understand how kinase function in the context of a whole cell assay. Cell-based assays provide a target in a more physiologically relevant environment, a natural substrate and information on permeability of drugs. Kinase therapeutics in oncology involves small molecule inhibitors as well as antibodies. Monoclonal antibodies in cancer target the extracellular domain of the kinase or ligand binding domain and a cell-based such as PathHunter™ assays provide a broad platform for such novel discoveries.

• No wash, single addition assay

• Utilizes full length, human protein

• Chemiluminescent assay, compatible with standard luminometers

• Whole, intact cells are used providing cell permeability information of compounds

• Measures ligand binding, receptor tyrosine phosphorylation adaptor protein binding and

• translocation

PathHunter™ Cell-Based Assay Portfolio

Technology Access

Targets Disease Relevance

Partner protein

Agonist Small MoleculeInhibitors/antibodies

DiscoveRx Part Number

TrkA

TrkB

TrkC

ErbB1

ErbB2-ErbB3

ErbB4

C-Met

PDGFRβ

FGFR4

INSR

IGFR1

EphB4

Flt3

CSF3R

PRLR - JAK1

PRLR - JAK2

FOXO3-AKT

For an updated target list, please visit www.discoverx.com

CNS

CNS

CNS

Cancer

Cancer

Cancer

Cancer

Cancer

Cancer

Diabetes

Diabetes

Prostrate Cancer

Cancer

Leukemia

Cancer

Cancer

Cancer

SHC1

SHC1

SHC1

PLCG2

GrB2

SHC1

GrB2

PLCG1

PLCG2

PLCG1

SHC1

SHC1

SHC1

SHC1

PLCG2

PLCG2

None

β-NGF

BDNF

NT3

EGF

Heregulin

NRG1

HGF

PDGF-AB

FGF-Basic

Insulin

IGF

Ephrin B2-FC

Flt-3 ligand

G-CSF

ProLactin

ProLactin

NA

93-0462C3

93-0463C3

93-0464C3

93-0681C3

93-0535C3

93-0465C3

93-0632C3

93-0469C3

93-0467C3

93-0466C3

93-0505C6

93-0468C3

93-0529C3

93-0564C3

93-0686C3

93-0687C3

93-0539C3

Cytosolic Tyrosine Kinase and Cytokine Receptors

Pathway Assays

Cytokine Receptor

Cytosolic

Phosphotyrosine Binding Domain

Jak1

FOXOPI3

InhibitionAKT

Inhibited

Nuclear TranslocationIncrease in signal

FOXO

FOXO

Light

PathHunter™ Cell-Based Kinase AssaysClonal cell lines expressing ProLink-tagged Receptor Tyrosine Kinase on the membrane and EA-SH2 fusion protein in the cytoplasm. Over 15 cell lines are now available. For more information, please visit www.discoverx.com/kinases.

Custom ProjectsUtilize DiscoveRx’s proprietary EFC technology to build your own functional cell-based kinase assays. Talk to our experts about your kinase targets, contact [email protected].

Custom Screening & Profiling ServicesSend your compound for simple profiling, specificity studies or selectivityprofiling against one or many receptor tyrosine kinase targets. For more information, please email [email protected].

Coming soon!

Coming soon!

score

score

score

score

score

score





















• translocation


Technology Access


Partner protein



TrkA

TrkB

TrkC

ErbB1

ErbB2-ErbB3

ErbB4

C-Met

PDGFRβ

FGFR4

INSR

IGFR1

EphB4

Flt3

CSF3R

PRLR - JAK1

PRLR - JAK2

FOXO3-AKT


CNS

CNS

CNS

Cancer

Cancer

Cancer

Cancer

Cancer

Cancer

Diabetes

Diabetes

Prostrate Cancer

Cancer

Leukemia

Cancer

Cancer

Cancer

SHC1

SHC1

SHC1

PLCG2

GrB2

SHC1

GrB2

PLCG1

PLCG2

PLCG1

SHC1

SHC1

SHC1

SHC1

PLCG2

PLCG2

None

β-NGF

BDNF

NT3

EGF

Heregulin

NRG1

HGF

PDGF-AB

FGF-Basic

Insulin

IGF

Ephrin B2-FC

Flt-3 ligand

G-CSF

ProLactin

ProLactin

NA

93-0462C3

93-0463C3

93-0464C3

93-0681C3

93-0535C3

93-0465C3

93-0632C3

93-0469C3

93-0467C3

93-0466C3

93-0505C6

93-0468C3

93-0529C3

93-0564C3

93-0686C3

93-0687C3

93-0539C3


Pathway Assays

Cytokine Receptor

Cytosolic


Jak1

FOXOPI3

InhibitionAKT

Inhibited


FOXO

FOXO

Light




Coming soon!

Coming soon!

score

score

score

score

score

score

PathHunter™ PathHunter Detection Reagents

Add compounds Add Read Luminescence

PathHunter™ Detection Reagents

Cytokine receptor

+ Jak1

Cytosolic tyrosine kinase

Phosphotyrosine binding domain

A. U2OS CSF3R-PK SHC1-EA JAK1

10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

G-CSF (g/mL)

RLU

B. U2OS SHC1 CSF3R JAK 1 with antagonists

10-15.0 10-12.5 10 -10.0 10 -7.5 10 -5.0 10 -2.5

0

1000

2000

3000

4000

5000

G-CSF

Staurosporine

Pyridone 6, P6, DBI (JAK Inhibitor 1)

Lestaurtinib

Compound

Mean

RLU

C. U2OS CSF3R-PK SHC1-EA JAK2

10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

8000

G-CSF (g/mL)

RLU

-12 -11 -10 -9 -8 -7 -6 -5 -40

25000

50000

75000

100000Wortmannin

LY294002

Akt inhibitor X

Inhibitor (M)

RLU

aa+ Ja

Jak1

10-11 10-10 10-9 10-8 10-7 10-6 10-52000

4000

6000

8000

10000

111.6

528

225

Blocking Ab (g/ml)

Mean

RLU

10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

1000

2000

3000

4000

β -NGF

anti-β-NGF

anti-beta-NGF/beta-NGF (g/mL)

RLU

10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

20

40

60

80

100

120

TrkA

TrkB

TrkC

Staurosporine (M)

No

rmali

zed

% A

ctiv

ity

A. Anti-Receptor Antibody B. Anti-Ligand Antibody C. Small Molecule Inhibitors

Target Specific PathHunter™ Receptor Tyrosine Kinase Assay Applications:Uncover Compound Specificity and Selectivity Information

Kinase drug discovery has historically focused on using purified kinases to identify ATP pocket binders. The availability of an almost complete panel of cell-based RTK assays allows one to profile such known kinase inhibitors to obtain selectivity and specificity information. The data below demonstrates that non-specific compound such as staurosporine inhibits almost all RTK/CTK assays whereas certain compounds maintain their selectivity.

Figure 1. PathHunter™ RTK and CTK Functional assays were used to profile the BioMol Kinase inhibitor library. After an overnight incubation, cells were treated with 10 μM of compound for 1 hour at 37°C and then with the respective ligands for 3 hours at 23 – 25°C. Percent inhibition of each compound against a specific RTK/CTK assay was calculated. This data was then used to generate a Heat chart that illustrates the value of profiling hits identified against all receptors in a specific family as well as other related kinases in a biologically relevant cell-based assay format.

Discover Novel Antibodies and Small Molecule Inhibitors

Figure 2. (A) A series of commercially available anti-receptor antibodies (Blocking antibodies) were tested with PathHunter™ U2OS cells expressing ErbB1 (93-0681C3). A dose dependent inhibition can be observed. (B) PathHunter™ U2OS cells expressing TrkA (93-0462C3) can also be used to profile anti-ligand antibodies. The data demonstrates agonist dose response and anti-ligand antibody’s inhibitory response. (C) PathHunter TrkA, TrkB and TrkC cells can be used to identify, screen or profile small molecule inhibitors.

PathHunter™ Cell-Based Pathway Assays Monitor Downstream Signaling

In this PathHunter assay, the U2OS cells have been engineered to express two complementing fragments of β-Gal within different cellular compartments. The small 42 AA enzyme fragment, ProLink is appended to FOXO-3. The larger enzyme fragment EA (Enzyme Acceptor) resides in the nucleus. Inhibition of PI3 or AKT allows the unphos-phorylated FOXO-3 to translocate to the nucleus facilitating complementation of the two enzyme fragments. This action results in the formation of fully complemented β-galactosidase enzyme, the activity of which is measured using chemiluminescent PathHunter Detection Reagents.

PathHunter™ Cell-Based Kinase Assay Protocol

• Single reagent addtion, no wash format • Standard 96 well and 384 well protocol; can be miniaturized to 1536 • Chemiluminescent, gain-of-signal assay • No special instrumentation required

These PathHunter cells are available as clonal cell lines. Bulk cell options are also available.

Get Closer to Your Target with PathHunter™ Receptor Tyrosine Kinase Assays

The PathHunter™ assay monitors the interaction of two proteins in a cell-based assay format using Enzyme Frag-ment Complementation (EFC). The cells have been engineered to express two complementing fragments of β-Gal appended to the proteins of interest. The PathHunter Receptor Tyrosine Kinase assay monitors the interaction of tyrosine phosphorylated receptors with SH2 containing proteins in a whole cell, homogeneous HTS-friendly assay. In this system, a small 42 AA enzyme fragment, ProLink (PK) is appended to the C-terminus of the receptor target. The SH2 protein is fused to the larger enzyme fragment, EA (Enzyme Acceptor). Activation of the receptor initiates dimerization and tyrosine phosphorylation of the receptor and subsequent binding to the SH2-EA fusion that forces complementation of the two enzyme fragments This action results in the formation of fully complementedβ-galactosidase enzyme, the activity of which is measured using chemiluminescent PathHunter Detection Reagents.

DiscoveRx offers the largest whole cell-based kinase portfolio with a 80% coverage of the Receptor tyrosine kinase family. PathHunter receptor tyrosine kinase assays offer a unique assay that measures ligand binding, phosphoryla-tion and interaction with downstream adaptor proteins. A full length receptor allows identification of non-ATP pocket binders, antibody therapeutics (Panel A), ligand binding inhibitors and dimerization inhibitors as well as small molecule inhibitors (Panel B).

Whole Cell Assay for Cytokine Receptors and Cytosolic Tyrosine Kinases Assays

Unlike receptor tyrosine kinases, the cytokine receptors lack kinase activity and are phosphorylated by cytosolic tyrosine kinases. To detect the phosphorylation status of cytokine receptors the ProLink is fused to the C-terminus of the target receptor, and co-expressed with a phosphotyrosine binding domain fused to the complementing fragment, EA. Upon activation and phosphorylation of the receptor by the cytosolic kinase, the SH2 domain binds to the phos-phorylated residues. Specificity for a specific kinase can be obtained by co-expression of specific tyrosine kinases.

Simple, One-Step, Whole Cell Assay for Cytokine Signaling and Screening

Figure 4. (A) Assays for Jak1 and Jak2 were developed using the G-CSFR-SHC1 PathHunter cell line. The G-CSFR is known to couple to both JaK1 and Jak2 kinases. Addition of the agonist, G-CSF, induces the activation of the expressed kinase resulting in phosphorylation of the target receptor, binding of SHC1 and complementation of the enzyme that is measured using Pathhunter reagents. (B) CSF3R cell line was incubated with and inhibitors such as staurosporine, Lestaurtinib and Pyridone 6, P6, DBI (JAK Inhibitor 1) and challenged with EC90 of their respective agonists (G-CSF, a known agonist). A dose dependent inhibition was observed indicating that the assay can be used to screen or profile inhibitors against the JAK molecule. (C) The assay measures CSF3R activation, phosphorylation by JAK2 and interaction between CSF3R, JAK2 and SHC1 proteins in a cell-based assay.

FOXO

PI3Inhibition

AKT Inhibited

Light


FOXO

FOXO

PPPPPPPPPPPPPP

Anti-ligandAnti-heterodimerization

antibody

KinaseInhibitor

Panel A

Panel B

Anti-receptor

Heterodimerization

Inhibitor

PD-9

8059

U-0

126

SB-2

0358

0H

-7H

-9St

auro

spor

ine

AG

-494

AG

-825

Lave

ndustin

ARG

-146

20Ty

rpho

stin

23Ty

rpho

stin

25Ty

rpho

stin

46Ty

rpho

stin

47Ty

rpho

stin

51Ty

rpho

stin

1Ty

rpho

stin

AG

128

8Ty

rpho

stin

AG

147

8Ty

rpho

stin

AG

129

5Ty

rpho

stin

9H

NM

PAPK

C-41

2Pi

ceat

anno

lPP

1A

G-4

90A

G-1

26A

G-3

70A

G-8

79LY

294

002

Wor

tman

nin

GF

1092

03X

Hyp

eric

inRo

31-

8220

Sphi

ngos

ine

H-8

9H

- 8

HA

-100

4H

A-1

077

HD

BAKN

-62

KN-9

3M

L-7

ML-

92-

Am

inop

urin

eN

9-Is

opro

pyl-o

lom

ouci

neO

lom

ouci

neis

o-O

lom

ouci

neRo

scov

itine

5-Io

dotu

berc

idin

LFM

-A13

SB-2

0219

0PP

2ZM

336

372

SU 4

312

AG

-129

6G

W 5

074

Palm

itoyl

-DL-

carn

itine

Cl

Rott

leri

nG

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Dai

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log

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DRB

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SP 6

0012

5In

diru

bin

Indi

rubi

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oxim

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2763

2Ke

npau

llone

Terr

eic

acid

Tric

irib

ine

BML-

257

SC-5

14BM

L-25

9A

pige

nin

BML-

265

(Erl

otini

b an

alog

)Ra

pam

ycin

TrkA

TrkC

ErbB2/3

c-MET

IGF1R

DDR2

PDGFRb

JAK1

0-20 20-4040

80-100

-60 60-80

PathHunter U2OS FOXO3 AKT Pathway Cells were plated in a 384-well plate and stimulated with the known PI3 and AKT inhibitors, PI3 and AKT inhibition facilitates FOXO translocation into the nucleus generating a positive gain of signal format in the PathHunter assay.

Features and Benefits

Assay Attribute Advantages

Full Length Receptor • Identify Anti-Receptor Antibodies

• Identify Anti-Ligand Antibodies

• Identify Activating Antibodies

• Identify Ligand Binding Inhibitors

• Identify Dimerization Inhibitors

• Detect Cytokine Receptor Phosphorylation

• Detect agonists or inhibitors for Cytokine Receptor Pathway Activation

• Monitor the role of cytosolic receptor kinases such as JAK1, JAK2 or JAK3 on cytokine receptor phosphorylation

• Identify JAK Inhibitors

Whole Cell Assay • Get cell permeability information

• Cellular processes are intact

Single Addition Assay

Assay measures ligand binding, phosphorylation and signal transduction. It measures the interaction between receptortyrosine kinases and adaptor proteins such as SHC, GrB2, PLCG1, PLCG2 or P85

• High-throughput friendly format


Novel Cell-Based Assay

• HTS-ready formatSimple, One-Step Assay

Benefits of PathHunter™ Pathway Assay

• Assay serves as PI3-kinase pathway indicator

• Allows for Screening of inhibitors for PI3-kinase and Akt

• A tool for studying molecules affecting FOXO3 nuclear translocation

score

score

score

score

score

score

Seedcells




Cytokine receptor

+ Jak1




10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

G-CSF (g/mL)

RLU


10-15.0 10-12.5 10 -10.0 10 -7.5 10 -5.0 10 -2.5

0

1000

2000

3000

4000

5000

G-CSF

Staurosporine


Lestaurtinib

Compound

Mean

RLU


10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

8000

G-CSF (g/mL)

RLU

-12 -11 -10 -9 -8 -7 -6 -5 -40

25000

50000

75000

100000Wortmannin

LY294002

Akt inhibitor X

Inhibitor (M)

RLU

aa+ Ja

Jak1

10-11 10-10 10-9 10-8 10-7 10-6 10-52000

4000

6000

8000

10000

111.6

528

225

Blocking Ab (g/ml)

Mean

RLU

10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

1000

2000

3000

4000

β -NGF

anti-β-NGF


RLU

10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

20

40

60

80

100

120

TrkA

TrkB

TrkC

Staurosporine (M)

No

rmali

zed

% A

ctiv

ity



















FOXO

PI3Inhibition

AKT Inhibited

Light


FOXO

FOXO

PPPPPPPPPPPPPP


antibody

KinaseInhibitor

Panel A

Panel B

Anti-receptor

Heterodimerization

Inhibitor

PD-9

8059

U-0

126

SB-2

0358

0H

-7H

-9St

auro

spor

ine

AG

-494

AG

-825

Lave

ndustin

ARG

-146

20Ty

rpho

stin

23Ty

rpho

stin

25Ty

rpho

stin

46Ty

rpho

stin

47Ty

rpho

stin

51Ty

rpho

stin

1Ty

rpho

stin

AG

128

8Ty

rpho

stin

AG

147

8Ty

rpho

stin

AG

129

5Ty

rpho

stin

9H

NM

PAPK

C-41

2Pi

ceat

anno

lPP

1A

G-4

90A

G-1

26A

G-3

70A

G-8

79LY

294

002

Wor

tman

nin

GF

1092

03X

Hyp

eric

inRo

31-

8220

Sphi

ngos

ine

H-8

9H

- 8

HA

-100

4H

A-1

077

HD

BAKN

-62

KN-9

3M

L-7

ML-

92-

Am

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9-Is

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5-Io

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LFM

-A13

SB-2

0219

0PP

2ZM

336

372

SU 4

312

AG

-129

6G

W 5

074

Palm

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BML-

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SC-5

14BM

L-25

9A

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nin

BML-

265

(Erl

otini

b an

alog

)Ra

pam

ycin

TrkA

TrkC

ErbB2/3

c-MET

IGF1R

DDR2

PDGFRb

JAK1

0-20 20-4040

80-100

-60 60-80

























score

score

score

score

score

score

Seedcells




Cytokine receptor

+ Jak1




10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

G-CSF (g/mL)

RLU


10-15.0 10-12.5 10 -10.0 10 -7.5 10 -5.0 10 -2.5

0

1000

2000

3000

4000

5000

G-CSF

Staurosporine


Lestaurtinib

Compound

Mean

RLU


10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

8000

G-CSF (g/mL)

RLU

-12 -11 -10 -9 -8 -7 -6 -5 -40

25000

50000

75000

100000Wortmannin

LY294002

Akt inhibitor X

Inhibitor (M)

RLU

aa+ Ja

Jak1

10-11 10-10 10-9 10-8 10-7 10-6 10-52000

4000

6000

8000

10000

111.6

528

225

Blocking Ab (g/ml)

Mean

RLU

10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

1000

2000

3000

4000

β -NGF

anti-β-NGF


RLU

10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

20

40

60

80

100

120

TrkA

TrkB

TrkC

Staurosporine (M)

No

rmali

zed

% A

ctiv

ity



















FOXO

PI3Inhibition

AKT Inhibited

Light


FOXO

FOXO

PPPPPPPPPPPPPP


antibody

KinaseInhibitor

Panel A

Panel B

Anti-receptor

Heterodimerization

Inhibitor

PD-9

8059

U-0

126

SB-2

0358

0H

-7H

-9St

auro

spor

ine

AG

-494

AG

-825

Lave

ndustin

ARG

-146

20Ty

rpho

stin

23Ty

rpho

stin

25Ty

rpho

stin

46Ty

rpho

stin

47Ty

rpho

stin

51Ty

rpho

stin

1Ty

rpho

stin

AG

128

8Ty

rpho

stin

AG

147

8Ty

rpho

stin

AG

129

5Ty

rpho

stin

9H

NM

PAPK

C-41

2Pi

ceat

anno

lPP

1A

G-4

90A

G-1

26A

G-3

70A

G-8

79LY

294

002

Wor

tman

nin

GF

1092

03X

Hyp

eric

inRo

31-

8220

Sphi

ngos

ine

H-8

9H

- 8

HA

-100

4H

A-1

077

HD

BAKN

-62

KN-9

3M

L-7

ML-

92-

Am

inop

urin

eN

9-Is

opro

pyl-o

lom

ouci

neO

lom

ouci

neis

o-O

lom

ouci

neRo

scov

itine

5-Io

dotu

berc

idin

LFM

-A13

SB-2

0219

0PP

2ZM

336

372

SU 4

312

AG

-129

6G

W 5

074

Palm

itoyl

-DL-

carn

itine

Cl

Rott

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nG

enis

tein

Dai

dzei

nEr

bstatin

ana

log

Que

rceti

n di

hydr

ate

SU14

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449

829

BAY

11-7

082

DRB

HBD

DE

SP 6

0012

5In

diru

bin

Indi

rubi

n-3'

-mon

oxim

eY-

2763

2Ke

npau

llone

Terr

eic

acid

Tric

irib

ine

BML-

257

SC-5

14BM

L-25

9A

pige

nin

BML-

265

(Erl

otini

b an

alog

)Ra

pam

ycin

TrkA

TrkC

ErbB2/3

c-MET

IGF1R

DDR2

PDGFRb

JAK1

0-20 20-4040

80-100

-60 60-80

























score

score

score

score

score

score

Seedcells




Cytokine receptor

+ Jak1




10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

G-CSF (g/mL)

RLU


10-15.0 10-12.5 10 -10.0 10 -7.5 10 -5.0 10 -2.5

0

1000

2000

3000

4000

5000

G-CSF

Staurosporine


Lestaurtinib

Compound

Mean

RLU


10-12 10-11 10-10 10-9 10-8 10-7 10-6

0

1000

2000

3000

4000

5000

6000

7000

8000

G-CSF (g/mL)

RLU

-12 -11 -10 -9 -8 -7 -6 -5 -40

25000

50000

75000

100000Wortmannin

LY294002

Akt inhibitor X

Inhibitor (M)

RLU

aa+ Ja

Jak1

10-11 10-10 10-9 10-8 10-7 10-6 10-52000

4000

6000

8000

10000

111.6

528

225

Blocking Ab (g/ml)

Mean

RLU

10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

1000

2000

3000

4000

β -NGF

anti-β-NGF


RLU

10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4

0

20

40

60

80

100

120

TrkA

TrkB

TrkC

Staurosporine (M)

No

rmali

zed

% A

ctiv

ity



















FOXO

PI3Inhibition

AKT Inhibited

Light


FOXO

FOXO

PPPPPPPPPPPPPP


antibody

KinaseInhibitor

Panel A

Panel B

Anti-receptor

Heterodimerization

Inhibitor

PD-9

8059

U-0

126

SB-2

0358

0H

-7H

-9St

auro

spor

ine

AG

-494

AG

-825

Lave

ndustin

ARG

-146

20Ty

rpho

stin

23Ty

rpho

stin

25Ty

rpho

stin

46Ty

rpho

stin

47Ty

rpho

stin

51Ty

rpho

stin

1Ty

rpho

stin

AG

128

8Ty

rpho

stin

AG

147

8Ty

rpho

stin

AG

129

5Ty

rpho

stin

9H

NM

PAPK

C-41

2Pi

ceat

anno

lPP

1A

G-4

90A

G-1

26A

G-3

70A

G-8

79LY

294

002

Wor

tman

nin

GF

1092

03X

Hyp

eric

inRo

31-

8220

Sphi

ngos

ine

H-8

9H

- 8

HA

-100

4H

A-1

077

HD

BAKN

-62

KN-9

3M

L-7

ML-

92-

Am

inop

urin

eN

9-Is

opro

pyl-o

lom

ouci

neO

lom

ouci

neis

o-O

lom

ouci

neRo

scov

itine

5-Io

dotu

berc

idin

LFM

-A13

SB-2

0219

0PP

2ZM

336

372

SU 4

312

AG

-129

6G

W 5

074

Palm

itoyl

-DL-

carn

itine

Cl

Rott

leri

nG

enis

tein

Dai

dzei

nEr

bstatin

ana

log

Que

rceti

n di

hydr

ate

SU14

98ZM

449

829

BAY

11-7

082

DRB

HBD

DE

SP 6

0012

5In

diru

bin

Indi

rubi

n-3'

-mon

oxim

eY-

2763

2Ke

npau

llone

Terr

eic

acid

Tric

irib

ine

BML-

257

SC-5

14BM

L-25

9A

pige

nin

BML-

265

(Erl

otini

b an

alog

)Ra

pam

ycin

TrkA

TrkC

ErbB2/3

c-MET

IGF1R

DDR2

PDGFRb

JAK1

0-20 20-4040

80-100

-60 60-80

























score

score

score

score

score

score

Seedcells





















• translocation


Technology Access


Partner protein



TrkA

TrkB

TrkC

ErbB1

ErbB2-ErbB3

ErbB4

C-Met

PDGFRβ

FGFR4

INSR

IGFR1

EphB4

Flt3

CSF3R

PRLR - JAK1

PRLR - JAK2

FOXO3-AKT


CNS

CNS

CNS

Cancer

Cancer

Cancer

Cancer

Cancer

Cancer

Diabetes

Diabetes

Prostrate Cancer

Cancer

Leukemia

Cancer

Cancer

Cancer

SHC1

SHC1

SHC1

PLCG2

GrB2

SHC1

GrB2

PLCG1

PLCG2

PLCG1

SHC1

SHC1

SHC1

SHC1

PLCG2

PLCG2

None

β-NGF

BDNF

NT3

EGF

Heregulin

NRG1

HGF

PDGF-AB

FGF-Basic

Insulin

IGF

Ephrin B2-FC

Flt-3 ligand

G-CSF

ProLactin

ProLactin

NA

93-0462C3

93-0463C3

93-0464C3

93-0681C3

93-0535C3

93-0465C3

93-0632C3

93-0469C3

93-0467C3

93-0466C3

93-0505C6

93-0468C3

93-0529C3

93-0564C3

93-0686C3

93-0687C3

93-0539C3


Pathway Assays

Cytokine Receptor

Cytosolic


Jak1

FOXOPI3

InhibitionAKT

Inhibited


FOXO

FOXO

Light




Coming soon!

Coming soon!

score

score

score

score

score

score





















• translocation


Technology Access


Partner protein



TrkA

TrkB

TrkC

ErbB1

ErbB2-ErbB3

ErbB4

C-Met

PDGFRβ

FGFR4

INSR

IGFR1

EphB4

Flt3

CSF3R

PRLR - JAK1

PRLR - JAK2

FOXO3-AKT


CNS

CNS

CNS

Cancer

Cancer

Cancer

Cancer

Cancer

Cancer

Diabetes

Diabetes

Prostrate Cancer

Cancer

Leukemia

Cancer

Cancer

Cancer

SHC1

SHC1

SHC1

PLCG2

GrB2

SHC1

GrB2

PLCG1

PLCG2

PLCG1

SHC1

SHC1

SHC1

SHC1

PLCG2

PLCG2

None

β-NGF

BDNF

NT3

EGF

Heregulin

NRG1

HGF

PDGF-AB

FGF-Basic

Insulin

IGF

Ephrin B2-FC

Flt-3 ligand

G-CSF

ProLactin

ProLactin

NA

93-0462C3

93-0463C3

93-0464C3

93-0681C3

93-0535C3

93-0465C3

93-0632C3

93-0469C3

93-0467C3

93-0466C3

93-0505C6

93-0468C3

93-0529C3

93-0564C3

93-0686C3

93-0687C3

93-0539C3


Pathway Assays

Cytokine Receptor

Cytosolic


Jak1

FOXOPI3

InhibitionAKT

Inhibited


FOXO

FOXO

Light




Coming soon!

Coming soon!

score

score

score

score

score

score

Documents

DRx brochure cell-based overview final v1 - DiscoverX Solutions for Drug Discovery · 2016. 10. 12. · Kinase targets have been extensively studied in biochemical assays using purified